Balanced salt solution preparation experiment
Balanced salt solution preparation experiment
Source of content: Tissue Culture and Molecular Cytology Techniques. (Beijing Publishing House)
Operation method
Experiment on the preparation of BSS
Principle
BSS is developed from Ringer physiological saline, mainly composed of inorganic salts and glucose.The inorganic ions in BSS are not only necessary for cell life, but also play an important role in maintaining osmotic pressure, buffering and regulating the pH of the solution.BSS contains a small amount of phenol red, which is used as an indicator for the change of the pH of the solution; it is yellow when the solution becomes acidic, purplish when it becomes alkaline, and peach when it is neutral. When the solution becomes acidic, it turns yellow, when it becomes alkaline, it turns purple, and when it becomes neutral, it turns peach, which makes it easy to observe the change of pH of culture solution. Move Hanks' liquid is used as an example of how to prepare it. Caveat 1. The main thing to avoid when preparing BSS is the precipitation of calcium and magnesium ions. For more product details, please visit Aladdin Scientific website.
1. Weigh the reagents accurately as shown in the table of ingredients; many reagents are hydrates containing water, which are different from those not containing water. Many reagents are hydrates containing water, which are different from those not containing water, and should be converted. 2;
2. Dissolve CaCl2 in 100 ml of water. 3;
3. dissolve the other components in 750 ml of water (note that the first reagent should be completely dissolved before dissolving the next component). (note that the first reagent should be completely dissolved before dissolving the next ingredient);
4. dissolve the phenol red with a few drops of 5.6 % NaHCO3 solution. 5. slowly pour 2 into 3 ml of water;
5. pour 2 slowly into 3, stirring occasionally to prevent precipitation. 6. add 4 to 5;
6. add 4 to 5;
7. transfer 6 into a volumetric flask and mix well with additional water;
8. Bottle, autoclave for 8 pounds 10 minutes, and store in refrigerator at 4℃.
According to the above method can also be formulated into 10 × concentration of the storage solution, in the storage solution with a little chloroform (2-3 ml) to prevent Chloroform can be volatilized during autoclaving, it will not affect the use; after adding chloroform, it should be mixed vigorously and stored at room temperature or 4 ℃. Storage at room temperature or 4 ℃. Dilute according to the proportion when using, filtered by filter paper and divided into bottles, packaged in autoclave, in sterile conditions, replace the sterile rubber stopper, placed in the refrigerator. After autoclaving the package, replace the sterile rubber stopper under sterile conditions and store in the refrigerator. If the liquid is turbid or precipitated, it should be reconstituted.